Coryneform bacteria, listeria and

erysipelothrix

Corynebacterium sp.
Listeria monocytogenes
Erysipelothrix rhusiopathiae
 Corynebacteria
• Significant Corynebacterium species
– C. xerosis
– C. pseudodiphtheriticum
– C. pseudotuberculosis
– C. jekeium
– C. ulcerans
• Rhodococcus equi
• Arcanobacterium haemolyticum
 Corynebacterium Species
• General characteristics
– Found as free-living saprophytes in fresh and salt water, in soil
and in the air
– Members of the usual flora of humans and animals
(often dismissed as contaminants)
– Often called “diphtheroids”
– Corynebacterium diphtheriae is the most significant pathogen
– Other species may cause infections in the immunocompromised
hosts
 Corynebacterium Species:
General Characteristics
• Morphology

– Gram-positive, non–spore-forming
rods
– Arrange in palisades:
“L-V” shape; “Chinese characters”
– Pleomorphic: “club-ends” or
coryneform
– Beaded, irregular staining
 C. diphtheriae: Agent of Diphtheria

• Toxigenic Corynebacterium diphtheriae

– Worldwide distribution but rare in places where vaccination
programs exist
• Exotoxin, Diphtheria toxin, as the virulence factor
– Not all C. diphtheriae strains produce toxin
– Toxin is produced by certain strains
– Toxin is antigenic
 Toxigenic Corynebacterium
diphtheriae
• Toxin consists of two fragments
– A: Active fragment
• Inhibits protein synthesis
• Leads to cell/tissue death
– B: Binding
• Binds to specific cell membrane receptors
• Mediates entry of fragment A into cytoplasm of host cell
 Clinical Forms of Diphtheria
• Respiratory
– Acquired by droplet spray or hand to mouth contact
– Non-immunized individuals are susceptible

• Non-respiratory
– Systemic
– Skin and cutaneous forms
 C. diphtheriae:
Causative Agent of Diphtheria
• Respiratory disease–diphtheria
– Incubation period–2 to 5 days

– Symptoms: sore throat, fever, malaise

– Toxin is produced locally, usually in the pharynx or tonsils

– Toxin causes tissue necrosis, can be absorbed to produce systemic

effects
– Forms a tough grey to white pseudomembrane which may cause
suffocation
 C. diphtheriae:
Causative Agent of Diphtheria
• C. diphtheriae pseudomembrane
– WBC + organism
 Clinical Infections:
Non-Respiratory Disease
• Systemic infections
– Toxin is absorbed in the blood stream and carried systemically

– Affects the kidneys, heart, and nervous system

– Death occurs due to cardiac failure

• Cutaneous form
– More prevalent in the tropics

– Infections occur at the site of minor abrasions

– Maybe superinfected with Streptococcus pyogenes and/or Staphylococcus

aureus
 Treatment

• Infected patients treated with anti-toxin and antibiotics

– Anti-toxin produced in horses
– Antibiotics have no effect on circulating toxin, but prevent spread
of the toxin
• Penicillin drug of choice
 Laboratory Diagnosis
• Microscopic morphology
– Gram-positive, non–spore-forming
rods, club-shaped, can be beaded
– Appear in palisades and give
"Chinese letter" arrangement
– Produce metachromatic granules or
“Babes’ Ernst” bodies (food
reserves) which stain more darkly
than remainder of organism

Corynebacterium diphtheriae
gram stain
 Laboratory Diagnosis:
Cultural Characteristics
• Loeffler's slant or Pai's slant—
Used to demonstrate
pleomorphism and metachromatic
granules ("Babes’ Ernst bodies“)

• Growth on Serum Tellurite or

modified Tinsdale exhibits brown
or grayish→ to black halos around
the colonies

Tellurite: tellurium dioxide (TeO2).

 Laboratory Diagnosis:
Cultural Characteristics

• Blood agar plae (BAP) 24-48 hours at 37oC

small, grey translucent colonies
• Small zone of - hemolysis also s een
 Laboratory Diagnosis
• Identification
– Confirm identification by fermentation reactions
(glucose +)
– Catalase positive
– Urease negative
– Non-motile
 Laboratory Diagnosis
• Toxigenicity testing
– Elek test
– Immunodiffusion test
• Organisms are streaked on
media with low Fe content
to maximize toxin
production.
• Identification of C.
diphtheriae does NOT
mean the patient has
dipheria. Must show the
isolate produces the toxin.

protease peptone agar + serum (horse or bovine)

1 and 4 positive
 C. diphtheriae
• Treatment: antitoxin
• Prevention: DPT immunization
 C. jekeium
• Clinical Infections
– Septicemia
– Meningitis
– Pulmonary disease

• Populations Affected
– Immunosuppressed
– IV drug users
 C. jekeium
Colony Morphology
• Isolation & identification
– BAP: 48-72 hours at 35oC in ambient air or 5%
CO2 small, gray-white colony, nonhemolytic
– Gram stain: pleomorphic, club-shaped gram
positive rod arranged in V forms or palisades
 C. jekeium
Lab Diagnosis
– Identification
• Nitrate reduction= negative
• Urea= negative
• Sucrose= negative
• Glucose= positive
 C. jekeium
• Susceptibility testing
– Exhibits resistance to multiple antibiotics
– Susceptible to vancomycin
Differentiating Characteristics of
Corynebacterium Species

Species Hemolysis Sucrose Nitrate Urease

fermentation reduction

C. diphtheriae V = + =
C. xerosis = + + =

C. pseudophtheriticum = = + +

C. pseudotuberculosis + V V +

R. equi = = + =

Corynebacterium = = = =
jekeium
 Listeria monocytogenes:
General Characteristics
• Gram-positive, non–spore-forming rods

• Only human pathogen in genus

• Widespread in nature

• Known to infect a wide variety of animals

• Human exposure is limited; direct or indirect

• Transient colonization occurs without disease

 Listeria monocytogenes:
Clinical Infections
• Adults
– Septicemia/meningitis in the compromised/elderly
– Mild flu-like syndrome in pregnant women could be fatal to fetus
– Ingestion of contaminated food
• Neonatal
– Early onset from intrauterine transmission results in sepsis; high
mortality rate
– Late onset manifests as meningitis; lower mortality rate
 Listeria monocytogenes:
• Virulence Factors
– Hemolysin ( Listeriolysin O)- damages macrophage
– Catalase
– Superoxide dismutase
– Phospholipid C
– P60 surface protein- induces phagocytosis thru
adhesion and penetration
 Laboratory Diagnosis:
L. monocytogenes
• Identification
– Microscopic morphology
• Gram Positive non–spore-
forming coccobacillary, pairs
or short chains

– Colony Morphology
• Grows well on blood agar;
colonies produce a narrow
zone of hemolysis similar to
Group B Streptococcus
• Small, round and translucent
 Laboratory Diagnosis:
L. monocytogenes
• Grows well at 0.5° C to 45° C
• Because of this temperature range, especially
the cooler end of the range, this organism
grows well in refrigerated products, such as
cream, cheese, deli meats, etc.
• Can sometimes be isolated after “cold
enrichment” (hold broth at 4° C for several
weeks and subculture)
 Laboratory Diagnosis:
L. monocytogenes
• Identification
– Catalase positive

– Motility:
• Motile at 25o C;
"umbrella" type →

• Tumbling motility in hanging

drop preparations (this can be
seen on Gram Stain Tutor at
www.medtraining.org)

“Umbrella” motility pattern (Left)

typical for L. monocytogenes
 Laboratory Diagnosis:
L. monocytogenes
• Identification
– CAMP test
• Produces a “block” type of
hemolysis in contrast to
“arrow”-shape produced by
Group B Streptococcus

CAMP test with

Listeria monocytogenes

Positive CAMP test for

Group B Streptococcus
 Differentiating Characteristics between
L. monocytogenes and Other Gram
Positive Bacteria
Hemolysis Motility Esculin Growth
Species Catalase At R. T. Hydrolysis 6.5% NaCl

L. monocytogenes + Beta + + +

Corynebacterium sp. + None, alpha =/+ = +/=

S. agalactiae = Beta = = =/+

Enterococcus sp. = None, alpha = + +

beta
 Erysipelothrix rhusiopathiae:
General characteristics
• Gram positive, non–spore-forming, pleomorphic rods (can
produce long filaments)

• Distributed in nature

• Can cause disease in animals (swine, turkey, sheep); swine is

the main reservoir

• Humans acquire the infection through occupational exposure,

such as cuts & scratches (fish handlers, animal products)
 Erysipelothrix rhusiopathiae:
Clinical Infections
• Erysipeloid
– Self-limiting localized infection at the site of inoculation
– Produces painful swelling, usually on the hands or fingers
– Heals within 3 to 4 weeks

• Endocarditis
– May occur in those who have had valve replacements
• Disseminated infections may occur, but rarely
 Laboratory Diagnosis:
Erysipelothrix rhusiopathiae
• Microscopic Morphology
– Pleomorphic, gram-positive thin rods that may form long
filaments, may be arranged singly, in short chains, or in a V
shape
 Laboratory Diagnosis:
Erysipelothrix rhusiopathiae
• Identification
– Catalase negative

– CO2 is required

– Distinguishing characteristic: Production of H2S on TSI

– Microaerophilic
– Nonmotile
– Test tube brush growth in semisolid motility media
 Laboratory Diagnosis:
Erysipelothrix rhusiopathiae
• Colony Morphology
– Grows on blood or chocolate agar
—colonies may appear gray or
translucent, pinpoint with alpha
hemolysis or nonhemolytic
 Treatment:
Erysipelothrix rhusiopathiae
• Penicillin, cephalosporin, erythromycin
Characteristics of Corynebacterium,
Listeria, and Erysipelothrix

Characteristics Corynebacterim Listeria Erysipelothrix

spp. monocytogenes rhusiopathiae

Hemolysis V + =
Discolored/
Catalase + + =
Growth at 4 o = + =
Motility at R.T. = + =
Esculin hydrolysis = + =
H2S (TSI butt) = = +